LOCUS       sequence-84588-e        4252 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4252)
  AUTHORS   Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske 
            M
  TITLE     A modular plasmid assembly kit for multigene expression, gene 
            silencing and silencing rescue in plants.
  JOURNAL   PLoS One. 2014 Feb 13;9(2):e88218. doi: 
            10.1371/journal.pone.0088218. eCollection 2014.
  PUBMED    24551083
REFERENCE   2  (bases 1 to 4252)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2014-02-13"; volume: "9(2):e88218. doi"; pages: " 
            10.1371/journal.pone.0088218. eCollection 2014"
COMMENT     Sequence Label: LIIc F 2-3
FEATURES             Location/Qualifiers
     source          1..4252
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     312..334
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     326..343
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     327..343
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     354..373
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        354..372
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    513..637
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(674..679,680..697,698..704)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             758..1417
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(824..843)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             1759..2064
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1943..1962
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2105..2229)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2429..2446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2539..3127)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2619..2638)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3298..4089,4090..4158))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3921..3940
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(4159..4252,1..11))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
       61 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
      121 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtggc cgctacaggg
      181 cgctcccatt cgccattcag gctgcgcaac tgttgggaag ggcgtttcgg tgcgggcctc
      241 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
      301 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgagcgcg acgtaatacg
      361 actcactata gggcgaattg gcggaaggcc gtcaaggcca cgtgtcttgt ccaggcgcgc
      421 catgagctct aaagcttcgc tcgagctgcg gccgcacgga tcctcgaatt cccaagctta
      481 tctcgacccg aagacaaggg cgcggtgaga ccacaagttt gtacaaaaaa gctgaacgag
      541 aaacgtaaaa tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta
      601 cataatactg taaaacacaa catatccagt cactatggcg gcccctcgag gggccgcatt
      661 aggcacccca ggctttacac tttatgcttc cggctcgtat aatgtgtgga ttttgagtta
      721 ggatccgtcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
      781 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
      841 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
      901 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
      961 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     1021 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     1081 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     1141 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     1201 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     1261 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     1321 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     1381 acaacagtac tgcgatgagt ggcagggcgg ggcgtaatct agaggatccg gcttactaaa
     1441 agccagataa cagtatgcgt atttgcgcgc tgatttttgc ggtataagaa tatatactga
     1501 tatgtatacc cgaagtatgt caaaaagagg tatgctatga agcagcgtat tacagtgaca
     1561 gttgacagcg acagctatca gttgctcaag gcatatatga tgtcaatatc tccggtctgg
     1621 taagcacaac catgcagaat gaagcccgtc gtctgcgtgc cgaacgctgg aaagcggaaa
     1681 atcaggaagg gatggctgag gtcgcccggt ttattgaaat gaacggctct tttgctgacg
     1741 agaacagggg ctggtgaaat gcagtttaag gtttacacct ataaaagaga gagccgttat
     1801 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
     1861 cccctggcca gtgcacgtct gctgtcagat aaagtccccc gtgaacttta cccggtggtg
     1921 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtttcc
     1981 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
     2041 aacctgatgt tctggggaat ataaatgtca ggctccctta tacacagcca gtctgcaggt
     2101 cgaccatagt gactggatat gttgtgtttt acagtattat gtagtctgtt ttttatgcaa
     2161 aatctaattt aatatattga tatttatatc attttacgtt tctcgttcag ctttcttgta
     2221 caaagtggtt ggggtctcat gtcattgttg tcttcggggt agcggccgct tggatcccag
     2281 aattctaggt acctcttaat taatggagca caagactggc ctcatgggcc ttccgctcac
     2341 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaacatggt catagctgtt
     2401 tccttgcgta ttgggcgctc tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     2461 gggtaaagcc tggggtgcct aatgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2521 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2581 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2641 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2701 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2761 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2821 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2881 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2941 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3001 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3061 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3121 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3181 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3241 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3301 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     3361 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     3421 tgctgcaatg ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca
     3481 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     3541 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     3601 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     3661 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     3721 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     3781 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     3841 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     3901 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     3961 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4021 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4081 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4141 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4201 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa ta
//
